Biological Cell Injection Using an Autonomous MicroRobotic System

Abstract

Recent advances in microbiology, such as cloning, demonstrate that increasingly complex micromanipulation strategies are required for manipulating individual biological cells. In this paper, we present a microrobotic system capable of conducting automatic embryo pronuclei DNA injection (cell injection). Conventionally, cell injection has been conducted manually, however, long training, low success rates from poor reproducibility in manual operations, and contamination all call for the elimination of direct human involvement. To automate cell injections, a microrobotic system is developed that is capable of performing automatic embryo pronuclei DNA injection autonomously through a hybrid visual servoing control scheme. A Hough transform is used to detect the nuclei of embryos. Sum-ofsquared-differences optical flow is used to track injection pipette motion, and auto focusing is implemented to determine the relative depth of subcellular structures. A hybrid control scheme is developed to fulfill the cell injection task. Upon the completion of injection, the DNA injected embryos are transferred into a pseudo-pregnant foster female mouse to reproduce transgenic mice for cancer studies. The experimental result shows that the injection success rate is 100%.

Quantifying growth mechanics of living, growing plant cells in situ using microrobotics

“…Despite the metrological limitations of the cantilever as a force sensor, few viable alternatives exist [13]. MEMS based force sensors have been used for multiple applications in biological research, such as for measuring forces on single heart cells [14], measuring the injection force on Drosophila embryos [15], studying cell mechanical response [16], characterizing fruit fly behavior and investigation of micromechanical properties of mouse zona pellucida and of soft hydrogel microcapsules [17][18][19]. These sensors are capable of measuring local properties of biological materials and microfabricated MEMS grippers have been demonstrated for quantifying global mechanical properties [20][21][22].…”

Vision-Based Haptic Feedback with Physically-Based Model for Telemanipulation

“…However, most cellular manipulation systems have primarily focused to date on visual information in conjunction with a dial-based console system. The operator needs extensive training to perform these tasks, and even an experienced operator can have low success rates and a poor reproducibility due to the nature of the tasks (Kallio & Kuncova, 2003;Sun & Nelson, 2002). The developed cell injection system is shown in Fig.…”

Evaluation of Telerobotic Shared Control Strategy for Efficient Single-Cell Manipulation

“…To address these problems, several single-cell microinjection systems have been proposed to improve the manipulation efficiency. Automated microinjection systems have been developed to remove human involvement from the injection process [10]- [16], where a visual servoing approach is usually used to control the position and force of a micromanipulator; however, it is challenging to create a fully automated manipulation system because microinjection is conducted under diverse and complex conditions such as varying cell size (from one micrometer to hundreds of micrometers), cell types (e.g., suspended or attached ones) and liquid environments. Fig.…”

“…Therefore, there are difficulties in the dexterous manipulation of cells with multiple degrees of freedom (DOF) and in target selection (e.g., cell nucleus or cytoplasm) in visual servoing [17], [18]- [20]. Teleoperated microinjection systems have been developed to provide haptic feedback during manipulation [3], [5], [7], [10], [15], [32]- [46].…”